Transcription mechanisms that contribute to HIV-1 latency

导致 HIV-1 潜伏期的转录机制

• 批准号: 8468555
• 负责人: Andrew J Henderson
• 金额: $ 41.25万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8468555
• 关键词: Anti-Retroviral Agents; Architecture; Biochemical; Biochemical Process; CD4 Positive T Lymphocytes; Cell Line; Cells; Chromatin; Complex; Coupled; Data; Event; Exhibits; Genes; Genetic Transcription; Goals; HIV; HIV Infections; HIV Long Terminal Repeat; HIV-1; In Vitro; Infection; Interruption; Libraries; Life; Long Terminal Repeats; Maintenance; Mass Spectrum Analysis; Molecular; Phenotype; Play; Positive Transcriptional Elongation Factor B; Properdin; Proteomics; Proviruses; RNA Polymerase II; Recruitment Activity; Regulation; Repression; Role; Source; System; T memory cell; T-Lymphocyte; Tissues; Transcription Elongation; Transcription Initiation; Transcription Repressor/Corepressor; Viral Proteins; Virus; cellular targeting; chromatin immunoprecipitation; chromatin remodeling; gene repression; insight; macrophage; negative elongation factor; novel; polarized cell; premature; promoter; public health relevance; purge; success; termination factor; transcription termination

DESCRIPTION (provided by applicant): HIV provirus is regulated at the level of transcription and involves changes in transcription initiation, chromatin organization and elongation. In particular, transcription elongation has been demonstrated to be a limiting step for HIV expression. We hypothesize that the coordinated control of RNA polymerase II processivity and premature transcription termination coupled with chromatin remodeling creates a key checkpoint that limits provirus transcription and directly impact HIV latency in different cellular and molecular contexts. We have shown that RNA polymerase II processivity is a check point for HIV transcription and that NELF, a factor that pauses RNA polymerase II, re-enforces repression of HIV proviruses by recruiting a termination factor Pcf11 and transcriptional corepressors to the HIV-1 long terminal repeat. Using a variety of biochemical approaches, including chromatin immunoprecipitation, RNAPII footprinting, in vitro transcription systems and mass spectrometry, we propose to determine the biochemical mechanisms and characterize the cellular factors that negatively regulate HIV transcription elongation. These studies will provide new insights into the establishment, maintenance and reversal of HIV latency. Understanding the regulation of HIV transcription elongation will provide novel cellular targets for controlling and purging HIV in different cellular reservoirs.

描述（由申请人提供）：HIV原病毒在转录水平受到调节，涉及转录起始、染色质组织和延伸的变化。特别是，转录延伸已被证明是 HIV 表达的限制步骤。我们假设 RNA 聚合酶 II 持续合成能力和过早转录终止的协调控制与染色质重塑相结合，创建了一个关键检查点，限制原病毒转录并直接影响不同细胞和分子背景下的 HIV 潜伏期。我们已经证明，RNA 聚合酶 II 的持续性是 HIV 转录的一个检查点，而 NELF（一种暂停 RNA 聚合酶 II 的因子）通过招募终止因子 Pcf11 和转录辅阻遏物到 HIV-1 长末端重复序列来重新强化对 HIV 前病毒的抑制。使用多种生化方法，包括染色质免疫沉淀、RNAPII 足迹、体外转录系统和质谱，我们建议确定生化机制并表征负调节 HIV 转录延伸的细胞因素。这些研究将为艾滋病毒潜伏期的建立、维持和逆转提供新的见解。了解 HIV 转录延伸的调节将为控制和清除不同细胞库中的 HIV 提供新的细胞靶点。